Tissue engineered vascular grafts (TEVGs) may provide vessel replacements when conventional prostheses are unavailable or fail. Researchers are taking multiple approaches toward fabricating viable TEVGs, including the development of designer scaffolds. A major limitation to rational TEVG scaffold design is the absence of correlative relationships between specific scaffold properties and resulting smooth muscle cell (SMC) behavior. We propose the development of novel PDMSstar-PEO hydrogels with tunable material properties to enable the systematic evaluation of scaffold property impact on SMC behavior and endpoint TEVG mechanical properties. Another novel element of this research design is our focus on the effects of scaffold properties on internal cellular signaling rather than on ECM synthesis alone. Specifically, we will examine the expression of an array of genes associated with the SRF pathway, a key regulatory pathway of SMC phenotype, in addition to ECM production and organization. A unique signature of ECM synthesis/organization plus gene expression will result and will serve as a predicative link between initial scaffold properties and endpoint TEVG mechanical properties, significantly advancing rational TEVG scaffold design. PUBLIC HEALTH RELEVANCE: The success of tissue engineering vascular grafts (TEVGs) has been limited largely due to insufficient long term mechanical properties, which results from inappropriate cell responses. In the proposed research we will prepare a library of novel hybrid scaffolds formed from hydrophobic and hydrophilic macromers whose properties may also be systematically tuned over a broad range. We will then explore the effects of systematic alterations in scaffold properties on smooth muscle cell behavior and long term TEVG mechanical properties toward the development of predictive relationships that can be used for rational TEVG scaffold design.